An improved direct intermicroscopic (LM leads to SEM leads to TEM) correlative procedure for the examination of mammalian skeletal muscle.

The procedure for direct intermicroscopic (LM leads to SEM leads to TEM) correlation of muscle was modified. The major modification was the use of cryostat sections which showed far better ultrastructural preservation than did paraffin sections. The use of an osmium-thiocarbohydrazide ligand binding procedure (OTOTO) did not noticeably improve the scanning electron microscopic (SEM) image, but did result in a subsequent transmission electron microscopic (TEM) image which showed increased contrast of the intracellular membranous components. Besides improving the preservation at the TEM level of examination, the techniques used to locate the same structure or area previously examined under the complementary modes of microscopy (i.e. LM and SEM), were also refined.